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12 pages, 1172 KiB

Open AccessCommunication

Retrospective Investigation of the Whole Genome of the Hypovirulent Listeria monocytogenes Strain of ST201, CC69, Lineage III, Isolated from a Piglet with Fatal Neurolisteriosis

by Sergey S. Zaitsev, Mariya A. Khizhnyakova and Valentina A. Feodorova

Microorganisms 2022, 10(7), 1442; https://doi.org/10.3390/microorganisms10071442 - 17 Jul 2022

Cited by 3 | Viewed by 1909

Abstract

Listeria monocytogenes (Lm), the causative agent for both human and animal listeriosis, is considered to be a rare but potentially fatal foodborne pathogen. While Lm strains associated with current cases of human listeriosis are now being intensely investigated, our knowledge of this microorganism [...] Read more.

Listeria monocytogenes (Lm), the causative agent for both human and animal listeriosis, is considered to be a rare but potentially fatal foodborne pathogen. While Lm strains associated with current cases of human listeriosis are now being intensely investigated, our knowledge of this microorganism which has caused listerial infection in the past is still extremely limited. The objective of this study was a retrospective whole-genome sequence analysis of the Lm collection strain, 4/52-1953, isolated in the middle of the 20th century from a piglet with listerial neuroinfection. The multi-locus sequence typing (MLST) analysis based on seven housekeeping genes (abcZ, bglA, cat, dapE, dat, ldh, and lhkA) showed that the Lm strain 4/52-1953 was assigned to the sequence type 201 (ST201), clonal complex 69 (CC69), and phylogenetic lineage III. The strain 4/52-1953, similarly to other ST201 strains, probably originated from the ST9, CC69 via ST157. At least eight different STs, ST69, ST72, ST130, ST136, ST148, ST469, ST769, and ST202, were identified as the descendants of the first generation and a single one, ST2290, was proved to be the descendant of the second generation. Among them there were strains either associated with some sporadic cases of human and animal listerial infection in the course of more than 60 years worldwide or isolated from food samples, fish and dairy products, or migratory birds. Phylogenetic analysis based on whole genomes of all the Lm strains available in the NCBI GenBank (n = 256) demonstrated that the strain 4/52-1953 belonged to minor Cluster I, represented by lineage III only, while two other major Clusters, II and III, were formed by lineages I and II. In the genome of the strain 4/52-1953, 41 virulence-associated genes, including the Listeria pathogenicity island 1 (LIPI-1), and LIPI-2 represented by two internalin genes, the inlA and inlB genes, and five genes related to antibiotic resistance, were found. These findings can help to make the emergence of both hyper- and hypovirulent variants, including those bearing antibiotic resistance genes, more visible and aid the aims of molecular epidemiology as well. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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12 pages, 2155 KiB

Open AccessArticle

Whole-Genome Sequencing Reveals Multiple Subpopulations of Dominant and Persistent Lineage I Isolates of Listeria monocytogenes in Two Meat Processing Facilities during 2011–2015

by Elton Burnett, Zuzana Kucerova, Molly Freeman, Sophia Kathariou, Jessica Chen and Monica Smikle

Microorganisms 2022, 10(5), 1070; https://doi.org/10.3390/microorganisms10051070 - 23 May 2022

Cited by 9 | Viewed by 2750

Abstract

Listeria monocytogenes is a foodborne pathogen with a highly clonal population structure comprising multiple phylogenetic sub-groups that can persist within food processing environments and contaminate food. The epidemiology of L. monocytogenes is well-described in some developed countries; however, little is known about the [...] Read more.

Listeria monocytogenes is a foodborne pathogen with a highly clonal population structure comprising multiple phylogenetic sub-groups that can persist within food processing environments and contaminate food. The epidemiology of L. monocytogenes is well-described in some developed countries; however, little is known about the prevalence and population structure of this pathogen in food and food processing environments located in less developed regions. The aim of this study was to determine the genetic characteristics and clonal relatedness of L. monocytogenes that were isolated from two Jamaican meat processing facilities. Of the 37 isolates collected between 2011 and 2015, only a single lineage II isolate was recovered (serotype 1/2c), and the remaining were lineage I isolates representing serotypes 4b, 1/2b, 3b, and two untypeable isolates. Pulsed-field gel electrophoresis (PFGE) delineated isolates into seven pulsotypes, and whole-genome sequencing (WGS) categorized most isolates within one of three clonal complexes (CC): CC2 (N = 12), CC5 (N = 11), and CC288 (N = 11). Isolates representing CC1 (N = 2) and CC9 (N = 1) were also recovered. Virulence-associated genes such as inlA and the LIPI-3 cluster were detected in multiple isolates, along with the stress survival islet cluster-1 (SSI-1), and benzalkonium (bcrABC) and cadmium (cad1, cad2, cad4) resistance cassettes. Multiple isolates that belong to the same CC and matching PFGE patterns were isolated from food and the environment from both facilities across multiple years, suggesting the presence of persistent strains of L. monocytogenes, and/or constant re-entry of the pathogens into the facilities from common sources. These findings highlight the ability of lineage I isolates of L. monocytogenes to colonize, persist, and predominate within two meat-producing environments, and underscores the need for robust surveillance strategies to monitor and mitigate against these important foodborne pathogens. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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17 pages, 1139 KiB

Open AccessArticle

Antimicrobial Activity of Ohelo Berry (Vaccinium calycinum) Juice against Listeria monocytogenes and Its Potential for Milk Preservation

by Biyu Wu, Xiaohan Liu, Stuart T. Nakamoto, Marisa Wall and Yong Li

Microorganisms 2022, 10(3), 548; https://doi.org/10.3390/microorganisms10030548 - 2 Mar 2022

Cited by 11 | Viewed by 2861

Abstract

Listeria monocytogenes is a foodborne pathogen and causes illnesses with a high mortality rate in susceptible populations. Several dairy-related outbreaks have been attributed to contamination by L. monocytogenes, which requires antimicrobial interventions to enhance the safety of these products. This study aimed [...] Read more.

Listeria monocytogenes is a foodborne pathogen and causes illnesses with a high mortality rate in susceptible populations. Several dairy-related outbreaks have been attributed to contamination by L. monocytogenes, which requires antimicrobial interventions to enhance the safety of these products. This study aimed to determine the antimicrobial activity of the ohelo berry (Vaccinium calycinum), a Hawaiian wild relative of cranberry, against L. monocytogenes in culture media and milk products. The effect of ohelo berry juice at its sub-inhibitory concentrations on the physicochemical properties, biofilm formation, and gene expression of L. monocytogenes was also investigated. The minimum inhibitory concentration of ohelo berry juice against L. monocytogenes was 12.5%. The sub-inhibitory concentration of ohelo berry juice (6.25%) significantly increased the auto-aggregation and decreased the hydrophobicity, swimming motility, swarming motility, and biofilm formation capability of L. monocytogenes. The relative expression of genes for motility (flaA), biofilm formation and disinfectant resistance (sigB), invasion (iap), listeriolysin (hly), and phospholipase (plcA) was significantly downregulated in L. monocytogenes treated by the 6.25% juice. L. monocytogenes was significantly inhibited in whole and skim milk supplemented with 50% ohelo berry juice, regardless of the fat content. These findings highlight the potential of ohelo berry as a natural preservative and functional food to prevent L. monocytogenes infection. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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15 pages, 989 KiB

Open AccessArticle

Distribution and Characteristics of Listeria spp. in Pigs and Pork Production Chains in Germany

by Verena Oswaldi, Stefanie Lüth, Janine Dzierzon, Diana Meemken, Stefan Schwarz, Andrea T. Feßler, Benjamin Félix and Susann Langforth

Microorganisms 2022, 10(3), 512; https://doi.org/10.3390/microorganisms10030512 - 26 Feb 2022

Cited by 13 | Viewed by 3260

Abstract

Listeria (L.) monocytogenes is a foodborne pathogen that can cause disease, mainly in elderly, pregnant or immunocompromised persons through consumption of contaminated food, including pork products. It is widespread in the environment and can also be found in asymptomatic carrier animals, [...] Read more.

Listeria (L.) monocytogenes is a foodborne pathogen that can cause disease, mainly in elderly, pregnant or immunocompromised persons through consumption of contaminated food, including pork products. It is widespread in the environment and can also be found in asymptomatic carrier animals, for example, in different tissues of pigs. To learn more about their nature, 16 Listeria spp. isolates found in tonsils and intestinal content of pigs and 13 isolates from the slaughterhouse environment were characterized using next-generation sequencing (NGS). A wide distribution of clonal complexes was observed in pigs, as well as in the pork production chain, suggesting multiple sources of entry. Hypervirulent clones were found in pig tonsils, showing the potential risk of pigs as source of isolates causing human disease. The presence of closely related isolates along the production chain suggests a cross-contamination in the slaughterhouse or recontamination from the same source, strengthening the importance of efficient cleaning and disinfection procedures. The phenotypical antimicrobial resistance status of L. monocytogenes isolates was examined via broth microdilution and revealed a low resistance level. Nevertheless, genotypical resistance data suggested multiple resistances in some non-pathogenic L. innocua isolates from pig samples, which might pose a risk of spreading resistances to pathogenic species. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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14 pages, 2244 KiB

Open AccessArticle

Modelling the Potential Risk of Infection Associated with Listeria monocytogenes in Irrigation Water and Agricultural Soil in Two District Municipalities in South Africa

by Chidozie Declan Iwu, Chinwe Juliana Iwu-Jaja, Rami Elhadi, Lucy Semerjian and Anthony Ifeanyin Okoh

Microorganisms 2022, 10(1), 181; https://doi.org/10.3390/microorganisms10010181 - 14 Jan 2022

Cited by 5 | Viewed by 2654

Abstract

Listeria monocytogenes (L. monocytogenes) is the etiologic agent of listeriosis which significantly affects immunocompromised individuals. The potential risk of infection attributed to L. monocytogenes in irrigation water and agricultural soil, which are key transmission pathways of microbial hazards to the human [...] Read more.

Listeria monocytogenes (L. monocytogenes) is the etiologic agent of listeriosis which significantly affects immunocompromised individuals. The potential risk of infection attributed to L. monocytogenes in irrigation water and agricultural soil, which are key transmission pathways of microbial hazards to the human population, was evaluated using the quantitative microbial risk assessment modelling. A Monte Carlo simulation with 10,000 iterations was used to characterize the risks. High counts of L. monocytogenes in irrigation water (mean: 11.96 × 10² CFU/100 mL; range: 0.00 to 56.67 × 10² CFU/100 mL) and agricultural soil samples (mean: 19.64 × 10² CFU/g; range: 1.33 × 10² to 62.33 × 10² CFU/g) were documented. Consequently, a high annual infection risk of 5.50 × 10⁻² (0.00 to 48.30 × 10⁻²), 54.50 × 10⁻² (9.10 × 10⁻³ to 1.00) and 70.50 × 10⁻² (3.60 × 10⁻² to 1.00) was observed for adults exposed to contaminated irrigation water, adults exposed to contaminated agricultural soil and children exposed to agricultural soil, respectively. This study, therefore, documents a huge public health threat attributed to the high probability of infection in humans exposed to L. monocytogenes in irrigation water and agricultural soil in Amathole and Chris Hani District Municipalities in the Eastern Cape province of South Africa. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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13 pages, 1522 KiB

Open AccessArticle

Whole-Genome Sequencing Characterization of Virulence Profiles of Listeria monocytogenes Food and Human Isolates and In Vitro Adhesion/Invasion Assessment

by Giuditta Fiorella Schiavano, Collins Njie Ateba, Annalisa Petruzzelli, Veronica Mele, Giulia Amagliani, Fabrizia Guidi, Mauro De Santi, Francesco Pomilio, Giuliana Blasi, Antonietta Gattuso, Stefania Di Lullo, Elena Rocchegiani and Giorgio Brandi

Microorganisms 2022, 10(1), 62; https://doi.org/10.3390/microorganisms10010062 - 28 Dec 2021

Cited by 23 | Viewed by 3016

Abstract

Listeria monocytogenes (Lm) is the causative agent of human listeriosis. Lm strains have different virulence potential. For this reason, we preliminarily characterised via Whole-Genome Sequencing (WGS) some Lm strains for their key genomic features and virulence-associated determinants, assigning the clonal complex [...] Read more.

Listeria monocytogenes (Lm) is the causative agent of human listeriosis. Lm strains have different virulence potential. For this reason, we preliminarily characterised via Whole-Genome Sequencing (WGS) some Lm strains for their key genomic features and virulence-associated determinants, assigning the clonal complex (CC). Moreover, the ability of the same strains to adhere to and invade human colon carcinoma cell line Caco-2, evaluating the possible correspondence with their genetic virulence profile, was also assessed. The clinical strains typed belonged to clonal complex (CC)1, CC31, and CC101 and showed a very low invasiveness. The Lm strains isolated from food were assigned to CC1, CC7, CC9, and CC121. All CC1 carried the hypervirulence pathogenicity island LIPI-3 in addition to LIPI-1. Premature stop codons in the inlA gene were found only in Lm of food origin belonging to CC9 and CC121. The presence of LIPI2_inlII was observed in all the CCs except CC1. The CC7 strain, belonging to an epidemic cluster, also carried the internalin genes inlG and inlL and showed the highest level of invasion. In contrast, the human CC31 strain lacked the lapB and vip genes and presented the lowest level of invasiveness. In Lm, the genetic determinants of hypo- or hypervirulence are not necessarily predictive of a cell adhesion and/or invasion ability in vitro. Moreover, since listeriosis results from the interplay between host and virulence features of the pathogen, even hypovirulent clones are able to cause infection in immunocompromised people. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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14 pages, 1062 KiB

Open AccessArticle

Behaviour of Listeria monocytogenes and Natural Microflora during the Manufacture of Riojano Chorizo (Spanish Dry Cured Sausage)

by Elena Gonzalez-Fandos, Maria Vazquez de Castro and Alba Martinez-Laorden

Microorganisms 2021, 9(9), 1963; https://doi.org/10.3390/microorganisms9091963 - 15 Sep 2021

Cited by 3 | Viewed by 2626

Abstract

Riojano chorizo is a dry cured sausage manufactured with traditional technologies without adding starter cultures at low temperatures. Its characteristics differ from other types of chorizo since sugars and nitrites are no added and processing temperatures are low- This work evaluates the behaviour [...] Read more.

Riojano chorizo is a dry cured sausage manufactured with traditional technologies without adding starter cultures at low temperatures. Its characteristics differ from other types of chorizo since sugars and nitrites are no added and processing temperatures are low- This work evaluates the behaviour of Listeria monocytogenes during the processing of inoculated Riojano chorizo as well as the natural microflora that can play a technological role or be of interest as indicators. The sausage mixture was inoculated with a cocktail of three selected strains of L. monocytogenes (CECT 932, CECT 934 and CECT 4032) (4 log₁₀ CFU/g) and after processed following the traditional production method. Samples were taken before inoculation, after inoculation, after stuffing (day 0) and on days 6, 13, 21 and 28 of processing. L. monocytogenes, mesophiles, Micrococcaceae, lactic acid bacteria, Enterobacteriaceae, S. aureus, sulfite-reducing clostridia and molds and yeast counts were evaluated. Furthermore, pH, water activity and humidity were determined. No growth of L mocytogenes was observed during the first 6 days, when the temperature of processing was 4 °C. The low temperature in the initial stages was a relevant hurdle to control L. monocytoegenes growth. A significant decrease (p ≤ 0.05) in L. monocytogenes counts was observed on day 13 compared to the initial counts. During drying (days 6 to 21) a reduction in this pathogen of 1.28 log CFU/g was observed. The low water activity below 0.92 on day 13 and 0.86 on day 21 seems to be critical for the reduction of L. monocytogenes. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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16 pages, 2915 KiB

Open AccessCommunication

Novel Sequence Types of Listeria monocytogenes of Different Origin Obtained in the Republic of Serbia

by Tatiana Yu. Bespalova, Tatiana V. Mikhaleva, Nadezhda Yu Meshcheryakova, Olga V. Kustikova, Kazimir Matovic, Marko Dmitrić, Sergey S. Zaitsev, Maria A. Khizhnyakova and Valentina A. Feodorova

Microorganisms 2021, 9(6), 1289; https://doi.org/10.3390/microorganisms9061289 - 12 Jun 2021

Cited by 8 | Viewed by 4169

Abstract

Listeria monocytogenes, the causative agent of listeriosis, is amongst the major food-borne pathogens in the world that affect mammal species, including humans. This microorganism has been associated with both sporadic episodes and large outbreaks of human listeriosis worldwide, with high mortality rates. [...] Read more.

Listeria monocytogenes, the causative agent of listeriosis, is amongst the major food-borne pathogens in the world that affect mammal species, including humans. This microorganism has been associated with both sporadic episodes and large outbreaks of human listeriosis worldwide, with high mortality rates. In this study, the main sequence types (STs) and clonal complexes (CCs) were investigated in all of the 13 L. monocytogenes strains originating from different sources in the Republic of Serbia in 2004–2019 and that were available in the BIGSdb-Lm database. We found at least 13 STs belonging to the phylogenetic lineages I and II. These strains were represented by ST1/ST3/ST9 of CC1/CC3/CC9, which were common in the majority of the European countries and worldwide, as well as by eight novel STs (ST1232/ST1233/ST1234/ST1235/ST1238/ST1236/ST1237/ST1242) of CC19/CC155/CC5/CC21/CC3/CC315/CC37, and the rare ST32 (clonal complex ST32) and ST734 (CC1), reported in the Republic of Serbia, the EU, for the first time. Our study confirmed the association of CC1 with cases of neuroinfection and abortions among small ruminants, and of CC3 and CC9 with food products of animal origin. The strains isolated in 2019 carried alleles of the internalin genes (inlA/inlB/inlC/inlE) characteristic of the most virulent strains from the hypervirulent CC1. These findings demonstrated the genetic relatedness between L. monocytogenes strains isolated in the Republic of Serbia and worldwide. Our study adds further information about the diversity of the L. monocytogenes genotypes of small ruminants and food products, as the strain distribution in these sources in Serbia had not previously been evaluated. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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15 pages, 754 KiB

Open AccessArticle

Low-Level Tolerance to Fluoroquinolone Antibiotic Ciprofloxacin in QAC-Adapted Subpopulations of Listeria monocytogenes

by Divya Kode, Ramakrishna Nannapaneni, Mohit Bansal, Sam Chang, Wen-Hsing Cheng, Chander S. Sharma and Aaron Kiess

Microorganisms 2021, 9(5), 1052; https://doi.org/10.3390/microorganisms9051052 - 13 May 2021

Cited by 8 | Viewed by 2737

Abstract

There was a development of low-level tolerance to fluoroquinolone antibiotic ciprofloxacin in Listeria monocytogenes after sublethal adaptation to quaternary ammonium compound (QAC). Using eight L. monocytogenes strains, we determined the changes in short-range MIC, growth rate, and survival for heterologous stress response to [...] Read more.

There was a development of low-level tolerance to fluoroquinolone antibiotic ciprofloxacin in Listeria monocytogenes after sublethal adaptation to quaternary ammonium compound (QAC). Using eight L. monocytogenes strains, we determined the changes in short-range MIC, growth rate, and survival for heterologous stress response to ciprofloxacin, after sublethal exposure to daily cycles of fixed or gradually increasing concentration of QAC. Three main findings were observed. (1) MIC increase—QAC-adapted subpopulations exhibited a significant increase in short-range MIC of ciprofloxacin, by 1.5 to 2.9 fold, as compared to non-adapted control for 4/8 strains (p < 0.05). (2) Growth rate increase—QAC-adapted subpopulations exhibited significant 2.1- to 6.8- fold increase in growth rate (OD₆₀₀ at 10 h) in ciprofloxacin-containing broth, as compared to non-adapted control for 5/8 strains (p < 0.05). (3) Survival increase—QAC-adapted subpopulations of L. monocytogenes yielded significantly higher survival in ciprofloxacin-containing agar by 2.2 to 4.3 log CFU/mL for 4/8 strains, as compared to non-adapted control (p ˂ 0.05). However, for other 4/8 strains of L. monocytogenes, there was no increase in survival of QAC-adapted subpopulations, as compared to non-adapted control in ciprofloxacin. These findings suggest the potential formation of low-level ciprofloxacin-tolerant subpopulations in some L. monocytogenes strains when exposed to residual QAC concentrations (where QAC might be used widely) and such cells if not inactivated might create food safety risk. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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21 pages, 3267 KiB

Open AccessEditor’s ChoiceArticle

Hypo- and Hyper-Virulent Listeria monocytogenes Clones Persisting in Two Different Food Processing Plants of Central Italy

by Fabrizia Guidi, Massimiliano Orsini, Alexandra Chiaverini, Marina Torresi, Patrizia Centorame, Vicdalia Aniela Acciari, Romolo Salini, Barbara Palombo, Giorgio Brandi, Giulia Amagliani, Giuditta Fiorella Schiavano, Francesca Romana Massacci, Stefano Fisichella, Marco Di Domenico, Massimo Ancora, Adriano Di Pasquale, Anna Duranti, Cesare Cammà, Francesco Pomilio and Giuliana Blasi

Microorganisms 2021, 9(2), 376; https://doi.org/10.3390/microorganisms9020376 - 13 Feb 2021

Cited by 28 | Viewed by 5409

Abstract

A total of 66 Listeria monocytogenes (Lm) isolated from 2013 to 2018 in a small-scale meat processing plant and a dairy facility of Central Italy were studied. Whole Genome Sequencing and bioinformatics analysis were used to assess the genetic relationships between [...] Read more.

A total of 66 Listeria monocytogenes (Lm) isolated from 2013 to 2018 in a small-scale meat processing plant and a dairy facility of Central Italy were studied. Whole Genome Sequencing and bioinformatics analysis were used to assess the genetic relationships between the strains and investigate persistence and virulence abilities. The biofilm forming-ability was assessed in vitro. Cluster analysis grouped the Lm from the meat plant into three main clusters: two of them, both belonging to CC9, persisted for years in the plant and one (CC121) was isolated in the last year of sampling. In the dairy facility, all the strains grouped in a CC2 four-year persistent cluster. All the studied strains carried multidrug efflux-pumps genetic determinants (sugE, mdrl, lde, norM, mepA). CC121 also harbored the Tn6188 specific for tolerance to Benzalkonium Chloride. Only CC9 and CC121 carried a Stress Survival Islet and presented high-level cadmium resistance genes (cadA1C1) carried by different plasmids. They showed a greater biofilm production when compared with CC2. All the CC2 carried a full-length inlA while CC9 and CC121 presented a Premature Stop Codon mutation correlated with less virulence. The hypo-virulent clones CC9 and CC121 appeared the most adapted to food-processing environments; however, even the hyper-virulent clone CC2 warningly persisted for a long time. The identification of the main mechanisms promoting Lm persistence in a specific food processing plant is important to provide recommendations to Food Business Operators (FBOs) in order to remove or reduce resident Lm. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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18 pages, 2826 KiB

Open AccessArticle

Population Structure of Non-ST6 Listeria monocytogenes Isolated in the Red Meat and Poultry Value Chain in South Africa

by Itumeleng Matle, Thendo Mafuna, Evelyn Madoroba, Khanyisile R. Mbatha, Kudakwashe Magwedere and Rian Pierneef

Microorganisms 2020, 8(8), 1152; https://doi.org/10.3390/microorganisms8081152 - 30 Jul 2020

Cited by 20 | Viewed by 3966

Abstract

Meat products have been implicated in many listeriosis outbreaks globally, however there is a dearth of information on the diversity of L. monocytogenes isolates circulating in food products in South Africa. The aim of this study was to investigate the population structure of [...] Read more.

Meat products have been implicated in many listeriosis outbreaks globally, however there is a dearth of information on the diversity of L. monocytogenes isolates circulating in food products in South Africa. The aim of this study was to investigate the population structure of L. monocytogenes isolated in the meat value chain within the South African market. Based on whole-genome sequence analysis, a total of 217 isolates were classified into two main lineage groupings namely lineages I (n = 97; 44.7%) and II (n = 120; 55.3%). The lineage groups were further differentiated into IIa (n = 95, 43.8%), IVb (n = 69, 31.8%), IIb (n = 28, 12.9%), and IIc (n = 25, 11.5%) sero-groups. The most abundant sequence types (STs) were ST204 (n = 32, 14.7%), ST2 (n = 30, 13.8%), ST1 (n = 25, 11.5%), ST9 (n = 24, 11.1%), and ST321 (n = 21, 9.7%). In addition, 14 clonal complex (CCs) were identified with over-representation of CC1, CC3, and CC121 in “Processed Meat-Beef”, “RTE-Poultry”, and “Raw-Lamb” meat categories, respectively. Listeria pathogenic islands were present in 7.4% (LIPI-1), 21.7% (LIPI-3), and 1.8% (LIPI-4) of the isolates. Mutation leading to premature stop codons was detected in inlA virulence genes across isolates identified as ST121 and ST321. The findings of this study demonstrated a high-level of genomic diversity among L. monocytogenes isolates recovered across the meat value chain control points in South Africa. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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Review

Jump to: Research

13 pages, 357 KiB

Open AccessReview

Nonhemolytic Listeria monocytogenes—Prevalence Rate, Reasons Underlying Atypical Phenotype, and Methods for Accurate Hemolysis Assessment

by Iwona Kawacka, Agnieszka Olejnik-Schmidt and Marcin Schmidt

Microorganisms 2022, 10(2), 483; https://doi.org/10.3390/microorganisms10020483 - 21 Feb 2022

Cited by 6 | Viewed by 2594

Abstract

Listeria monocytogenes is a foodborne pathogen that typically presents β-hemolytic activity. However, there are literature reports indicating that L. monocytogenes strains are sometimes nonhemolytic or their zones of hemolysis are perceivable only after removal of the colonies from the agar plate. Nonhemolytic L. [...] Read more.

Listeria monocytogenes is a foodborne pathogen that typically presents β-hemolytic activity. However, there are literature reports indicating that L. monocytogenes strains are sometimes nonhemolytic or their zones of hemolysis are perceivable only after removal of the colonies from the agar plate. Nonhemolytic L. monocytogenes are most commonly encountered in food products, but some have also been detected in clinical samples. Usually, atypical bacteria of this species belong to serotype 1/2a. Mutations of the prfA gene sequence are the most common reason for changed phenotype, and mutations of the hly gene are the second most common cause. There are also reports that the methodology used for detecting hemolysis may influence the results. Sheep or horse blood, although most commonly used in modern studies, may not allow for the production of clear hemolytic zones on blood agar, whereas other types of blood (guinea pig, rabbit, piglet, and human) are more suitable according to some studies. Furthermore, the standard blood agar plate technique is less sensitive than its modifications such as bilayer or top-layer (overlay) techniques. The microplate technique (employing erythrocyte suspensions) is probably the most informative when assessing listerial hemolysis and is the least susceptible to subjective interpretation. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

22 pages, 2194 KiB

Open AccessReview

Application of Whole Genome Sequencing to Aid in Deciphering the Persistence Potential of Listeria monocytogenes in Food Production Environments

by Natalia Unrath, Evonne McCabe, Guerrino Macori and Séamus Fanning

Microorganisms 2021, 9(9), 1856; https://doi.org/10.3390/microorganisms9091856 - 31 Aug 2021

Cited by 21 | Viewed by 5503

Abstract

Listeria monocytogenes is the etiological agent of listeriosis, a foodborne illness associated with high hospitalizations and mortality rates. This bacterium can persist in food associated environments for years with isolates being increasingly linked to outbreaks. This review presents a discussion of genomes of [...] Read more.

Listeria monocytogenes is the etiological agent of listeriosis, a foodborne illness associated with high hospitalizations and mortality rates. This bacterium can persist in food associated environments for years with isolates being increasingly linked to outbreaks. This review presents a discussion of genomes of Listeria monocytogenes which are commonly regarded as persisters within food production environments, as well as genes which are involved in mechanisms aiding this phenotype. Although criteria for the detection of persistence remain undefined, the advent of whole genome sequencing (WGS) and the development of bioinformatic tools have revolutionized the ability to find closely related strains. These advancements will facilitate the identification of mechanisms responsible for persistence among indistinguishable genomes. In turn, this will lead to improved assessments of the importance of biofilm formation, adaptation to stressful conditions and tolerance to sterilizers in relation to the persistence of this bacterium, all of which have been previously associated with this phenotype. Despite much research being published around the topic of persistence, more insights are required to further elucidate the nature of true persistence and its implications for public health. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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20 pages, 834 KiB

Open AccessReview

Listeria monocytogenes Cold Shock Proteins: Small Proteins with A Huge Impact

by Francis Muchaamba, Roger Stephan and Taurai Tasara

Microorganisms 2021, 9(5), 1061; https://doi.org/10.3390/microorganisms9051061 - 14 May 2021

Cited by 27 | Viewed by 4646

Abstract

Listeria monocytogenes has evolved an extensive array of mechanisms for coping with stress and adapting to changing environmental conditions, ensuring its virulence phenotype expression. For this reason, L. monocytogenes has been identified as a significant food safety and public health concern. Among these [...] Read more.

Listeria monocytogenes has evolved an extensive array of mechanisms for coping with stress and adapting to changing environmental conditions, ensuring its virulence phenotype expression. For this reason, L. monocytogenes has been identified as a significant food safety and public health concern. Among these adaptation systems are cold shock proteins (Csps), which facilitate rapid response to stress exposure. L. monocytogenes has three highly conserved csp genes, namely, cspA, cspB, and cspD. Using a series of csp deletion mutants, it has been shown that L. monocytogenes Csps are important for biofilm formation, motility, cold, osmotic, desiccation, and oxidative stress tolerance. Moreover, they are involved in overall virulence by impacting the expression of virulence-associated phenotypes, such as hemolysis and cell invasion. It is postulated that during stress exposure, Csps function to counteract harmful effects of stress, thereby preserving cell functions, such as DNA replication, transcription and translation, ensuring survival and growth of the cell. Interestingly, it seems that Csps might suppress tolerance to some stresses as their removal resulted in increased tolerance to stresses, such as desiccation for some strains. Differences in csp roles among strains from different genetic backgrounds are apparent for desiccation tolerance and biofilm production. Additionally, hierarchical trends for the different Csps and functional redundancies were observed on their influences on stress tolerance and virulence. Overall current data suggest that Csps have a wider role in bacteria physiology than previously assumed. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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11 pages, 269 KiB

Open AccessReview

Detection and Potential Virulence of Viable but Non-Culturable (VBNC) Listeria monocytogenes: A Review

by Nathan E. Wideman, James D. Oliver, Philip Glen Crandall and Nathan A. Jarvis

Microorganisms 2021, 9(1), 194; https://doi.org/10.3390/microorganisms9010194 - 19 Jan 2021

Cited by 45 | Viewed by 6085

Abstract

The detection, enumeration, and virulence potential of viable but non-culturable (VBNC) pathogens continues to be a topic of discussion. While there is a lack of definitive evidence that VBNC Listeria monocytogenes (Lm) pose a public health risk, recent studies suggest that Lm in [...] Read more.

The detection, enumeration, and virulence potential of viable but non-culturable (VBNC) pathogens continues to be a topic of discussion. While there is a lack of definitive evidence that VBNC Listeria monocytogenes (Lm) pose a public health risk, recent studies suggest that Lm in its VBNC state remains virulent. VBNC bacteria cannot be enumerated by traditional plating methods, so the results from routine Lm testing may not demonstrate a sample’s true hazard to public health. We suggest that supplementing routine Lm testing methods with methods designed to enumerate VBNC cells may more accurately represent the true level of risk. This review summarizes five methods for enumerating VNBC Lm: Live/Dead BacLight^TM staining, ethidium monoazide and propidium monoazide-stained real-time polymerase chain reaction (EMA- and PMA-PCR), direct viable count (DVC), 5-cyano-2,3-ditolyl tetrazolium chloride-4′,6-diamidino-2-phenylindole (CTC-DAPI) double staining, and carboxy-fluorescein diacetate (CDFA) staining. Of these five supplementary methods, the Live/Dead BacLight^TM staining and CFDA-DVC staining currently appear to be the most accurate for VBNC Lm enumeration. In addition, the impact of the VBNC state on the virulence of Lm is reviewed. Widespread use of these supplemental methods would provide supporting data to identify the conditions under which Lm can revert from its VBNC state into an actively multiplying state and help identify the environmental triggers that can cause Lm to become virulent. Highlights: Rationale for testing for all viable Listeria (Lm) is presented. Routine environmental sampling and plating methods may miss viable Lm cells. An overview and comparison of available VBNC testing methods is given. There is a need for resuscitation techniques to recover Lm from VBNC. A review of testing results for post VBNC virulence is compared Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

19 pages, 415 KiB

Open AccessReview

Listeria monocytogenes Biofilms in the Food Industry: Is the Current Hygiene Program Sufficient to Combat the Persistence of the Pathogen?

by Tina Mazaheri, Brayan R. H. Cervantes-Huamán, Maria Bermúdez-Capdevila, Carolina Ripolles-Avila and José Juan Rodríguez-Jerez

Microorganisms 2021, 9(1), 181; https://doi.org/10.3390/microorganisms9010181 - 15 Jan 2021

Cited by 84 | Viewed by 8389

Abstract

Biofilms contain microbial cells which are protected by a self-produced matrix and they firmly attach themselves to many different food industry surfaces. Due to this protection, microorganisms within biofilms are much more difficult to eradicate and therefore to control than suspended cells. A [...] Read more.

Biofilms contain microbial cells which are protected by a self-produced matrix and they firmly attach themselves to many different food industry surfaces. Due to this protection, microorganisms within biofilms are much more difficult to eradicate and therefore to control than suspended cells. A bacterium that tends to produce these structures and persist in food processing plants is Listeria monocytogenes. To this effect, many attempts have been made to develop control strategies to be applied in the food industry, although there seems to be no clear direction on how to manage the risk the bacteria poses. There is no standardized protocol that is applied equally to all food sectors, so the strategies for the control of this pathogen depend on the type of surface, the nature of the product, the conditions of the food industry environment, and indeed the budget. The food industry performs different preventive and corrective measures on possible L. monocytogenes-contaminated surfaces. However, a critical evaluation of the sanitization methods applied must be performed to discern whether the treatment can be effective in the long-term. This review will focus on currently used strategies to eliminate biofilms and control their formation in processing facilities in different food sectors (i.e., dairy, meat, fish, chilled vegetables, and ready-to-eat products). The technologies employed for their control will be exemplified and discussed with the objective of understanding how L. monocytogenes can be improved through food safety management systems. Full article

(This article belongs to the Special Issue An Update on Listeria monocytogenes)

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